Pharmaceutical compositions

ABSTRACT

A composition including a synergistic combination of a vitamin D3 or a derivative or precursor thereof and hyaluronic acid or derivate thereof encapsulated in a lipid based colloidal carrier system (preferably lipid based vesicles such as liposomes, niosomes, tranferosomes) and topical formulations thereof, as well as their use in the prevention and/or treatment of inflamed skin and mucous membrane, especially in the prevention and/or treatment of skin photodamage, in particular in the prevention and/or treatment of skin erythema (skin inflammation) and actinic keratosis, as well non melanoma skin cancer.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention is directed towards new compositions comprising asynergistic combination of a vitamin D or a derivative or precursorthereof and hyaluronic acid or derivate thereof encapsulated in a lipidbased colloidal carrier system (preferably lipid based vesicles such asliposomes, niosomes, tranferosomes) and topical formulations thereof, aswell as their use in the prevention and/or treatment of inflamed skinand mucous membrane, especially in the prevention and/or treatment ofskin photodamage, in particular in the prevention and/or treatment ofskin erythema (skin inflammation) and actinic keratosis, as well nonmelanoma skin cancer.

Discussion of Related Art

Skin disorders according to ICD-10 (International Classification ofDiseases, Version 2016) include (a) group of conditions in which theskin becomes inflamed, forms blisters, and becomes crusty, thick, andscaly (including eczema causing burning and itching, occurring over along period of time), (b) any type of skin inflammation, (c) aninflammatory process affecting the skin (with signs of red rash,itching, and blister formation), e.g. contact dermatitis, atopicdermatitis, seborrheic dermatitis and psoriasis, and (d) pruriticpapulovesicular dermatitis occurring as a reaction to many endogenousand exogenous agents. Photo damage of the skin (according to ICD-10version 2016) is characterized as a skin disorder due to radiation ofultraviolet A (UVA) and ultraviolet B (UVB) with the following majorsymptoms: skin atrophy, skin dyspigmentation (patches/spots),photodermatitis (erythema: inflamed, reddened skin), telangiectasia,(couperose) and actinic keratosis. UVB with a wavelength of 280-315 nmprovides the energy the skin needs to make vitamin D3, but is also aprimary mutagen that penetrates through the epidermal layer of the skin,resulting in DNA mutations, potentially leading to skin cancer (nonmelanoma skin cancer (NMSC) and melanoma). These mutations may beclinically related to specific signs of photodamage such as increasingin elastin and collagen defects resulting in skin atrophy. UVA with awavelength of 315-400 nm is able to penetrate deeper into the skin ascompared to UVB rays and thus may damage both the epidermal and dermallayers. With constant UVA exposure, the size of the dermis layer will bereduced, causing an atrophy of the skin. Potential damages includedilated or broken blood vessels, causing telangiectasia (couperose) orindirect damages to cellular DNA as well as lipids and proteins of theskin barrier through the generation of reactive oxygen species (ROS),which are cytotoxic. Both UVA and UVB exposure can also lead toinflammation and vasodilation, which is clinically manifested astelangiectasia, (couperose) and photodermatitis (erythema resulting ininflamed and reddened skin), Dyspigmentation (patches/spots) and otherskin disorders [see e.g. 1, 2, 3].

In inflamed skin tissue high concentrations of reactive oxygen species(ROS) like nitric oxide are present. Nitric oxide (NO) reacts furtherwith oxygen (O₂) to peroxynitrite (ONOO⁻). Peroxynitrite and itsdegradation from the reaction with CO₂ (NO₂ ⁻ and CO₃ ⁻) are highlycytotoxic throughout the oxidation of lipids, proteins and DNA in theepidermis.

Various approaches have been suggested to counter skin inflammationssuch as photodamage and its effects, including the uses of vitamins, forexample vitamin D3 and its derivatives and precursors, as well ashyaluronic acid and the like.

Vitamin D is a group of fat-soluble vitamins with vitamin D₃ (orcholecalciferol) and vitamin D₂ (or ergocalciferol) being the mostimportant representatives in humans. Typically, vitamin D3 is obtainedby photolysis of 7-dehydrocholesterol (or 7-DHC, found prominently inthe stratum spinosum and stratum basale of the upper layer of the skinat about 25-50 ug/cm²) by UVB [see e.g. 4-7, FIG. 1] to obtain theprecursor previtamin D3, which is then thermally isomerized to givevitamin D3. It is known that the vitamin D3 production throughout theskin decreases dramatically with aging (up to 75 wt % at the age of 70).Vitamin D3 and its precursors and derivatives are biologically veryactive. For example, the precursor 7-DHC has the capability to bind thereactive oxigenes species NO and thus avoids overproduction of cytotocicperoxynitrite in the upper parts of the skin, preventing the viciouscircle (circulus vitiosus) of skin inflammation with cellular skindamages [FIGS. 2, 3]. Vitamin D3 plays a role in many processes,including bone mineralization, bone growth and bone remodelling,modulation of cell growth, neuromuscular and immune function, andothers. It has also been proposed that certain vitamin D3 analogues(e.g. 25-hydroxy-vitamin-D3 or calcidiol, 1, 25-dihydroxyvitamin-D3 orcalcitriol, calcipotriol; see e.g. 8, 9, 10, FIG. 3) can be usedtopically to treat skin conditions, including psoriasis [see e.g. 11,12, 13, 14]. In addition, recent studies using genetically modifiedmice, which exhibit altered mineral homeostasis due to a high vitamin D3activity, showed features of premature aging that include retardedgrowth, osteoporosis, atherosclerosis, ectopic calcification,immunological deficiency, skin and general organ atrophy. This and otherfindings suggests that serum calcidiol might be associated with anincreased risk of aging-related chronic diseases including cancer.Vitamin D is also involved in rebuilding the skin barrier [see e.g. 15,FIG. 2], sustaining immune defence against microorganisms and protectinga healthy microflora [see e.g. 16], Vitamin D3 also reduce inflammation,supporting of the skin and is involved in the wound healing [see e.g.17-20], and protecting the skin from photo damage [see e.g. 21-25, FIGS.2, 3]. It is presumed that throughout the local production of vitamin Din the skin together with the skin browning is the most importantmechanism of skin protection against photo damage [see e.g. 26].

Hyaluronic acid (HA) is a linear polysaccharide with repeatingdisaccharide units composed of glucuronic acid and N-acetyl glucosamineand is one of the major matrix substances in which cells and fibrousconstituents of the matrix such as collagen and elastin are embedded[see e.g. 27, 28, FIG. 2]. HA has an enormously high water bindingcapacity [see e.g. 29] and contributes largely to the maintenance of theextracellular space and to control tissue hydration working as ahumectant [see e.g. 30]. It is known, that crosslinking HA polymerchains transform the HA solution into a gel. Crosslinker molecules bindindividual HA polymer chains to create a network, which manifestsmacroscopically as a gel mass. It has been suggested that HA plays apivotal role in tissue regeneration [see e.g. 31, 32, FIG. 2].

However, despite the numerous formulations that are commercialized forskin treatments, there is still a high need for a formulation that iscapable to prevent and/or treat the common symptoms of skininflammation, especially photodamage, namely the affected skin and evenmore to prevent skin damages caused by sun exposure, especially by UVradiation, with greater effectiveness.

Applicants have now found that vitamin D3 or a derivative or precursorthereof, preferably a precursor such as 7-dehydrocholesterol (7-DHC) ora derivative thereof can be formulated in combination with HA as astabilized colloid. This colloidial carrier system shows a synergisticeffect in the treatment and prevention of inflamed skin, especiallyphotodamage of the skin, such as skin atrophy, skin dyspigmentation(patches/spots), photodermatitis (erythema: inflamed and reddened skin),telangiectasia, (couperose) and UV prevention to avoid skin erythema andactinic keratosis [FIG. 2].

Thus applicants provide a new composition comprising vitamin D3 or aderivative or precursor thereof, preferably a precursor such as 7-DHC ora derivative thereof and HA or derivatives, optionally in combinationwith additional excipients, encapsulated in a lipid based colloidalcarrier system (preferably lipid based vesicles such as liposomes,niosomes, tranferosomes) to allow the penetration and localized deliveryof stabilized vitamin D3 or a derivative or precursor thereof,preferably a precursor such as 7-DHC or a derivative thereof into theupper layers of the skin.

The two active substances combined topically are able to act in asynergistic manner directly in the upper layers of the skin on thedominant disorders of inflamed skin, especially in photodamage, inparticular on the indication according ICD-10 of sun and especially UVradiation exposed and damaged skin. Thus the new composition of theinvention is able to overcome drawbacks of prior art. The additionalseven excipients will act synergistic with the two active substances andallow an optimal efficacy to prevent and/or treat the described skindisorders. The described composition (and formulations thereof) willprovide a new approach to prevent and/or treat most disorders ofinflamed skin, sun and especially UV radiation damaged skin [FIGS. 2,3].

SUMMARY OF THE INVENTION

In a first aspect, the invention is directed towards a new composition(also referred to as composition of the invention) comprising asynergistic combination of at least one vitamin D3 or a derivative orprecursor thereof, preferably a precursor such as 7-DHC or a derivativethereof and at least one HA or derivative thereof, encapsulated in alipid based colloidal carrier system (to allow for optimal skinpenetration and stabilization of the vitamin D3 or a precursor orderivative thereof) and suitable formulations thereof. By encapsulationin the oil (or lipid) phase the vitamin D3 or a precursor or derivativethereof (and in particular 7-DHC or a derivative thereof) is stabilizedand undesired reactions (such as oxidation or other degradationreactions) are eliminated. The lipid based colloidal carrier systemallows vitamin D3 (and inactive vitamin D3 precursors such as 7-DHC or aderivative thereof) to penetrate into the upper layers of the skin,where exerts its activity (after being converted into the active vitaminD3 upon UVA and UVB exposure).

In a preferred embodiment the vitamin D3 is a vitamin D3 precursor suchas 7-DHC or a derivative thereof, and the composition of the inventioncomprises a vitamin D precursor such as 7-DHC or a derivative thereof,in combination with HA or a derivative thereof encapsulated in a lipidbased colloidal carrier system. The vitamin D (and in particular thevitamin D3 precursor such as 7-DHC or a derivative thereof) ispreferably present in the colloidal carrier system at a finalconcentration of 0.01 to 0.5 wt %.

In other embodiments the composition further comprises one or more,preferably 1, 2, 3, 4, 5, 6, 7 further components selected from avitamin A (preferably retinyl palmitate), at least one vitamin B, avitamin C (preferably L-ascorbic acid) and a vitamin E (preferablytocopheryl acetate). Preferably, the composition further comprises oneor more, most preferably all, of (i) retinyl palmitate (vitamin A), (ii)riboflavin (vitamin B2), (iii) niacinamide (vitamin B3), (iv)dexpanthenol (Provitamin B5), (v) folic acid (vitamin B9), (vi)L-ascorbic acid (vitamin C) and (vii) tocopheryl acetate (vitamin E).

In another specific embodiment, the lipid based colloidal carrier systemis a liposomal carrier system (preferably a lipid based vesicle such asliposome, niosome, tranferosome) composed of at least one phospholipidand at least one fatty acid. Preferably the lipid based colloidalcarrier system comprises one or more of e.g. lecithin, linolenic acid,linoleic acid, phosphatidylcholine and caprylic/capric triglyceride.

In a preferred embodiment, the lipid based colloidal carrier (e.g. alipid based vesicle such as liposome, niosome, tranferosome) comprisesthe synergistic combination of 7-DHC and HA in combination with the twolipophilic agents retinyl palmitate (Vitamin A), tocopheryl acetate(Vitamin E), and the five hydrophilic agents riboflavin (Vitamin B2),niacinamide (Vitamin B3), dexpanthenol (Provitamin B5), folic acid(Vitamin B9), L-ascorbic acid (Vitamin C).

The lipophilic agents are encapsulated within the bilayer ormultilamellar system, whereas the hydrophilic agents are encapsulated inthe aqueous phase. Thus, in one embodiment the composition of theinvention is obtained by (i) encapsulating the vitamin D3 (and inparticular vitamin D3 precursor such as 7-DHC or a derivative thereof)in the oil (or lipid) phase of the lipid based colloidal carrier systemat room temperatures, and (ii) separately preparing HA in the aqueousphase. The compositions are obtained by emulsification of the waterphase with the oil (or lipid) phase (by mixing or spontaneousintegration at room temperature). Preferably, the particles will have adiameter of 10-500 nm, more preferably 10-300 nm, most preferably 20-150nm.

In a further aspect, the composition of the invention is in form ofvarious formulations suitable for topical or transdermal and mucosaadministration. These topical formulations contain the pharmaceuticalcomposition of the invention, as well as further auxiliary agents, suchas buffering agents, preserving agents and the like. Typicalformulations include hydrogels, liogels, hydrolotions, lipolotions,cremes, ointments, and the like.

In a further aspect, the invention is directed towards the use of thecomposition of the invention (and topical formulations thereof) in theprevention and/or treatment of skin photo damage symptoms, in particularin the prevention and/or treatment of skin atrophy, skin dyspigmentation(patches/spots), photodermatitis (erythema: inflamated and reddenedskin), telangiectasia, (couperose), prevention of photodermatitis(erythema: inflammation of the skin), actinic keratosis and skin UVA andUVB protection.

BRIEF DESCRIPTION OF SEVERAL VIEWS OF THE DRAWINGS

FIG. 1: Adequate concentrations of the inactive precursor 7-DHC istransported in form of a lipid-based colloid according to the inventionin the upper layers of the skin, where it is converted to active vitaminD3 upon UVA and UVB exposure.

FIG. 2: 7-DHC in the oil (or lipid) phase and hyaluronic acid (HA) inthe water phase are integrated into the colloidal carrier system, whichallows the penetration into the upper layers of the skin. With theactivation and synthesis of vitamin D3 from 7-DHC the pathway of skindamages by sunlight UVA/UVB can be blocked. Inactive 7-DHC is protectedfrom oxidative processes with the encapsulation into the carrier systemand further being only activated at the target into the upper layers ofthe skin having a potent function eliminating cytotoxic ROS asperoxynitrates, resulting from the oxidative processes of nitrates. Thisscavenger function of in excess produced NO's allows the protection ofskin damages from UVA/UVB.

FIG. 3: 7-DHC pathway of potent anti-inflammation activity in the upperlayers of the skin after being activated to vitamin D3.

DETAILED DESCRIPTION OF THE INVENTION

In a first aspect, the invention is directed towards a new composition,hereinafter also called composition of the invention, comprising asynergistic combination of at least one vitamin D3 or a precursor orderivative thereof, preferably a precursor such as 7-dehydrocholesterol(7-DHC) or a derivative thereof, and at least one HA or derivativethereof, encapsulated in a lipid based colloidal carrier system(preferably lipid based vesicles such as liposomes, niosomes,tranferosomes), and suitable topical formulations thereof. Alldefinitions and embodiments specified hereinafter apply to thecompositions (and topical formulations) of the invention and usesthereof (unless specified otherwise). The term “topical” as used hereinrefers to administration to any part of the skin and mucous membranes,including ocular mucous membranes. The term “photodamage” as used hereinrefers to ICD-10 definition 2016 and is characterized as a skin disorderdue to sun exposure and to radiation of UVA and UVB. The term“synergistic” when used in relation to the compositions of the presentinvention means that the therapeutic effect of the combination of agentsis greater than the sum of the effects of the individual agents in thecombination.

The term “vitamin D” as used herein refers to any of the antirachiticforms known in the art to be suitable for nutritional use such asvitamin D₁, vitamin D₂, vitamin D₃, vitamin D₄, vitamin D₅, vitamin D₆,and vitamin D₇. Preferred is“vitamin D3”, which as used herein refers tovitamin D3 as well as a precursor of vitamin D3, such as 7-DHC(provitamin D3) or a derivative thereof, or a derivative of vitamin D3,such as 25-hydroxyvitamin D3, or 1a, 25-dihydroxyvitamin D3, including,1a-hydroxyvitamin D3, that activates the vitamin D receptor or that canbe metabolically converted in a human to a compound that activates thevitamin D receptor. Preferred is 7-DHC. The vitamin D, preferably thevitamin D3 and its precursor 7-DHC, is used at a concentration of 10,000IU-50,000 IE and 0.01-4 wt %. Preferable concentration of 0.01 to 3 wt%, more preferably of 0.01 to 0.75 wt %, most preferably 0.01 to 0.5 wt% of the total weight of the composition according to the presentinvention.

The term “hyaluronic acid” (also known as hyaluronan, hyaluronate, orHA) as used herein refers to an unsulphated glycosaminoglycan composedof repeating disaccharide units of N-acetylglucosamine (GlcNAc) andglucuronic acid (GlcUA) linked together by alternating beta-1,4 andbeta-1,3 glycosidic bonds. The term “hyaluronic acid” or “HA” as used inthe present application refers to HA or salts of HA, such as the sodium,potassium, magnesium and calcium salts, among others. The term“hyaluronic acid” or “HA” includes both natural and synthetic formulasand combinations of these natural and synthetic formulas including saltforms thereof. HA and its various molecular size fractions and therespective salts thereof have been used as medicaments, especially intreatment of arthropathies, as an auxiliary and/or substitute agent fornatural organs and tissues, especially in ophthalmology and cosmeticsurgery, and as agents in cosmetic preparations. Products of HA havealso been developed for use in orthopaedics, rheumatology, anddermatology. High molecular weight (MW) fractions of HA having anaverage MW of about 1 to about 1.5 MDa are well known for providingexcellent moisturizing properties in cosmetic compositions such aslotions and creams. Very low MW fractions of HA have been reported tohave a higher ability to penetrate the skin barrier. In preferredembodiments, a crosslinking agent (e.g. 1,4-butanediol diglycidal ether(BDDE) and the like) can be used to bind HA polymer chains to eachother, transforming liquid HA solutions into gels. Thus, in a specificembodiment, HA is in form of a gel obtained by crosslinking the HApolymer chains (through the primary hydroxyl site (—CH₂OH) and/orsecondary hydroxyl sites (—CHOH) within the HA monomeric unit), with lowmolecular crosslinked HA showing a high water retention capacity intothe skin. HA for use in the present invention is preferably of low MW,e.g. 4 kDa to 50 kDa, combined with higher MW up to 200′000 kDa.Typically, the HA is used at a concentration of 0.01 to 8 wt % (or 80mg/ml), preferably of 0.01 to 5 wt % (or 50 mg/ml), more preferably of0.01 to 4 wt % (or 40 mg/ml), most preferably 0.01 to 3 wt % (or 30mg/ml). The most preferable concentration of total HA is 3 wt %,preferably as a mixture of lowest MW HA of 4-5 kDa, low to medium ormedium molecular HA of 40-50 kDa and high MW HA of 50′000-200′000 kDa(wt %). Preferably the ratio of lowest molecular HA of 4-5 kDa to mediummolecular HA of 40-50 kDa to high MW HA of 50′000-200′000 kDa is(1-10):(0.1-2):(0.1-2), preferably (2-6):(0.5-1.5):(0.5-1.5), mostpreferably about 4:about 1:about 1 (or about equal wt % of medium andhighMW HA). Thus, most preferred is as a 3 wt % HA mixture of 2 wt % oflowest molecular HA of 4-5 kDa, 0.5 wt % of low to medium or mediummolecular HA of 40-50 kDa and 0.5 wt % of high MW HA of 50′000-200′000kDa.

Together with a vitamin D, such as vitamin D3 such as 7-DHC, HA has asynergistic effect on the hydration of the epidermis and also on theimmune protective effect [see e.g. 33, 34, 35, 36 FIGS. 2, 3]. HAtogether with a vitamin D3 such as 7-DHC have a synergisticphysico-chemical mode of action on photo damaged skin.

Thus, in a preferred embodiment the composition of the inventioncomprises 7-DHC in the oil phase of the colloidal carrier system and HAas active substances in the water phase of the colloidal carrier system.The two phases are obtained separately and then combined to form a lipidbased colloidal carrier system.

It was further found that 7-DHC also serves as additional activator ofthe further components (hereinafter also referred to as auxiliaryagents), which are particularly effective in the prevention and/ortreatment of photodamage. In particular, it was found that compositionsfurther comprising one or more components selected from a vitamin A(preferably retinyl palmitate), at least one vitamin B, a vitamin C(preferably L-ascorbic acid) and a vitamin E (preferably tocopherylacetate) achieve an effective prevention and/or treatment of photodamage[see FIGS. 1, 2, 3].

Thus in specific embodiments the composition of the invention furthercomprises one or more, preferably 1, 2, 3, 4, 5, 6, or 7 furthercomponents selected from a vitamin A (preferably retinyl palmitate), atleast one vitamin B, a vitamin C (preferably L-ascorbic acid) and avitamin E (preferably tocopheryl acetate).

The term “vitamin A” as used herein refers to retinol, retinal, retinoicacid, and several provitamin A carotenoids (most notably beta-carotene),preferably the major form retinyl palmitate. Vitamin A and particularlyretinyl palmitate absorbs light in the short-wavelength UVA range,having a photoprotective effect in the skin. It was found that retinylpalmitate showed an ad on effect together with 7-DHC with regard toabsorbing short-wavelength UVA range, the down regulation of NF-κB andtherefore on the UV-induced inflammation of the skin [see e.g. 36, 37].Retinyl palmitate diffuses into the skin, where it is partiallyhydrolyzed to retinol, penetrates into the stratum corneum, epidermis,and dermis and acts as a UV filter by absorbing UV radiation in therange between 300-350 nm theregy supporting the effects of thecompositions of the invention. Typically, retinyl palmitate is used at aconcentration of 0.01 wt % up to 2 wt %, preferably 0.01 wt % to 0.5 wt%, more preferably 0.01 to 0.2 wt %, most preferably 0.01 to 0.1 wt %.

The term “vitamin B” as used herein refers a class of water-soluble,chemically distinct vitamins including thiamine (B1), riboflavin (B2),niacin (B3), pantothenic acid (B5), pyridoxine (B6), folate (B7) andvarious cobalamins (B12). In one embodiment the term “vitamin B” as usedherein refers to riboflavin (B2). Typically, riboflavin is used at aconcentration of 0.01 wt % to 2 wt %, preferably 0.01 wt % to 0.2 wt %,more preferably 0.01 wt % to 0.1 wt %, most preferably 0.01 wt % to0.0.05 wt % of the total weight of the composition according to thepresent invention. In another embodiment, the term “vitamin B” as usedherein refers to niacinamide. Niacinamide, an amide of niacin (B3), is ahydrophilic endogenous substance, which has the potential to act as anantioxidant, can improve epidermal barrier function, decrease skinhyperpigmentation, reduce skin atrophy, decrease redness/blotchiness,and improve skin elasticity [54, 55]. Niacinamide shows a synergisticeffect with HA in rebuilding the structural and functional integrity ofthe epidermal barrier function and as humectant of the epidermis [FIG.2]. Niacinamide controls the NFκB-mediated transcription of signallingmolecules by inhibiting the nuclear poly (ADP-ribose) polymerase-1(PARP-1). Additionally niacinamide will have an added on effect onNFκB-mediated transcription with 7-DHC and vitamin A (particularlyretinyl palmitate) [see e.g. 38, 40]. Typically, niacinamide (vitaminB3) is used at a concentration of 0.5 wt % up to 5 wt %, preferably upto 4 wt %, more preferably up to 3 wt %, most preferably 3 wt % of thetotal weight of the composition according to the present invention.

In another embodiment, the term “vitamin B” as used herein refers todexpanthenol (provitamin B5). Topical dexpanthenol acts like a humectantand the activity may be based on the hygroscopic properties ofdexpanthenol. Dexpanthenol additionally shows protective effects againstskin irritation [see e.g. 39. Dexpanthenol significantly accelerates thewound healing process in children post-tonsillectomy intervention [seee.g. 36]. Typically, dexpanthenol (vitamin B5) is used at aconcentration of 0.5 wt % up to 5 wt %, preferably of to 3 wt %, morepreferably of 2.5 wt %, most preferably 1 wt % of the total weight ofthe composition according to the present invention.

In a further embodiment the term “vitamin B” as used herein refers tofolic acid (B9). Folic acid is essential for DNA synthesis, repair andmethylation, in particular nucleotide biosynthesis and remethylation ofhomocysteine. Folic acid is essential for cellular DNA, RNA production,and is known for its use in the prevention of neural tube defects (NTDs)and serious birth defects and the treatment of anaemia caused by folicacid deficiency. Folic acid also shows in vitro and in vivo incombination with creatine a significant acceleration of the epidermalskin regeneration [see e.g. 41] and thus, can promote a synergisticeffect together with 7-DHC covering the UV-induced cell damages andinflammation [FIG. 2]. Typically, folic acid (Vitamin B9) is used at aconcentration of 0.01 wt % up to 0.2 wt %, preferably of to 0.07 wt %,more preferably of 0.05 wt %, most preferably 0.02 wt % of the totalweight of the composition according to the present invention.

The term “vitamin C” as used herein refers to L-ascorbic acid, which isused as a supplement to treat and prevent scurvy and erythema of theskin [see e.g. 42, 43]. Scurvy leads to the formation of brown spots onthe skin, spongy gums, and bleeding from all mucous membranes.L-ascorbic acid acts as an electron donor for different essentialenzymes in the skin, which are required for the hydroxylation of prolineand lysine in the synthesis of collagen [see e.g. 44, 45, 46 and thesynthesis of carnitine, which is essential for the transport of fattyacids into mitochondria for ATP generation in the dermal cells [see e.g.47, 48]. Ascorbate also acts as an antioxidant, protecting againstoxidative stress [see e.g. 49] and is a powerful reducing agent capableof rapidly scavenging a number of reactive oxygen species (ROS) andthus, can promote a synergistic effect together with 7-DHC. Typically,L-ascorbic acid is used at a concentration of 0.1 wt % up to 10 wt %,preferably of to 5 wt %, more preferably of 2 wt %, most preferably 3 wt% of the total weight of the composition according to the presentinvention.

The term “vitamin E” as used herein refers to compounds known astocopherols and tocotreienols, preferably tocopheryl acetate. Tocopherylacetate can penetrate the skin to the living cells, where about 5 wt %is converted to free tocopherol. Tocopheryl acetate has shownantioxidant activities and acts as a peroxyl radical scavenger,disabling the production of damaging free radicals in tissues [see e.g.50] and thus can promote a synergistic effect together with 7-DHC.Typically, tocopheryl acetate is used at a concentration of 0.1 wt % to5 wt %, preferably 0.1 wt % to 5 wt %, more preferably 0.1 wt % to 3 wt%, most preferably 0.1 wt % to 2 wt % of the total weight of thecomposition according to the present invention.

Thus, in preferred embodiments, the composition of the inventioncomprising 7-DHC and HA further comprises one or more componentsselected from retinyl palmitate, riboflavin, niacinamide, dexpanthenol,folic acid, L-ascorbic acid and tocopheryl acetate, encapsulated in alipid based colloidal carrier system. In specific embodiments thecomposition of the invention comprises 7-DHC and HA as well as acombination of components as follows:

a) Retinyl palmitate and riboflavin or retinyl palmitate and niacinamideor retinyl palmitate and dexpanthenol or retinyl palmitate and folicacid or retinyl palmitate and L-ascorbic acid or retinyl palmitate andtocopheryl acetate.

b) Riboflavin and niacinamide or riboflavin and dexpanthenol orriboflavin and (v) folic acid or riboflavin and L-ascorbic acid orriboflavin and tocopheryl acetate.

c) Niacinamide and dexpanthenol or niacinamide and folic acid orniacinamide and L-ascorbic acid or niacinamide and tocopheryl acetate(vitamin E).

d) Dexpanthenol and folic acid or dexpanthenol and L-ascorbic acid ordexpanthenol and tocopheryl acetate.

e) Folic acid and L-ascorbic acid or folic acid and tocopheryl acetate.

f) L-ascorbic acid and tocopheryl acetate.

Most preferably, the compositions (and topical formulations of theinvention comprising 7-DHC and HA further comprise the componentsretinyl palmitate, riboflavin, niacinamide, dexpanthenol, folic acid,L-ascorbic acid and tocopheryl acetate encapsulated in a lipid basedcolloidal carrier system. The most preferable concentrations and rangesof concentrations are described as follows in Table 1.

TABLE 1 Active Conc. Range Preferred conc. Ingredients* wt % wt %CAS-No. 7-DHC 0.01-5   0.15  434-16-2 HA 0.01-5   3 (or 30 mg/ml)9004-61-9 Retinyl palmitate 0.01-0.5 0.500 79-81-2 Riboflavin 0.01-0.20.100 83-88-5 Niacinamide 0.5-4  4.000 98-92-0 Dexpanthenol  0.5-2.52.500 81-13-0 Folic acid 0.01-0.2 0.050 59-30-3 L-ascorbic acid  0.1-103.000 50-81-7 Tocopheryl 0.1-5  2.000 7695-91-2 Acetate *The activeingredients are incorporated in a colloidal carrier system according tothe invention (typically: water: >50 wt %, e.g. 50-75 wt %; carriersystem 10 wt %; additional oily ingredient, conservants, buffers,filters, 5-25 wt %)

The term “lipid based colloidal carrier (system)” (or “colloid”) as usedherein refers well known particulate carrier systems, preferablyspherical vesicles having at least one lipid bilayer. Typical colloidalcarriers include liposomes, niosomes, transferosomes, micelles,nanoparticles, microemulsions and others, preferably liposomes,niosomes, transferosomes, most preferably liposomes. Depending on theirsize and number of bilayers, the lipid based colloidal carrier systemisin form of: (a) multilamellar vesicles (MLV), (b) large unilamellarvesicles (LUV), (c) small unilamellar vesicles (SUV), (d) multivesiclevesicles (MW), oligolamellar vesicles (OLV). The preferred particle sizeranges from 10-500 nm, preferably 10 to 300 nm, more preferably 20-150nm.

In specific embodiments, the colloids are based on natural and/orsynthetic phospholipids and compose typically 10 wt % of theformulation. Typically used phospholipids include fatty acids having aphosphate-containing polar endgroup which is hydrophilic and thussoluble in water, and a hydrophobic end group, which is soluble in fatsjoined together by a glycerol molecule (e.g. glycerophospholipids) orsphingosine molecule (e.g. phosphosphingolipids).

In some embodiments, the phospholipids used in the colloidal carriersystem include one or more of phosphatidylcholine,lysophosphotidylcholine, hydrogenated phospholipids, and unsaturatedphospholipids. Examples of glycerophospholipids include phosphatidicacid (phosphatidate) (PA), phosphatidylethanolamine (cephalin) (PE),phosphatidylcholine (lecithin) (PC), Phosphatidylserine (PS), andPhosphoinositides, which further include phosphatidylinositol (PI),phosphatidylinositol phosphate (PIP), phosphatidylinositol bisphosphate(PIP2), and phosphatidylinositol triphosphate (PIP3). Examples ofphosphosphingolipids include ceramide phosphorylcholine (sphingomyelin)(SPH), ceramide phosphorylethanolamine (sphingomyelin) (Cer-PE), andceramide phosphorylglycerol. The colloidal carrier system of theinvention may further comprise fatty acids such as omega-3, omega-6 andomega-9 fatty acids. Preferred examples used in the present inventionare lecithin, sphingomyeline, phosphatidylcholine, linoleic acid,linolenic acid, caprylic acid, capric acid, Lupinus albus seed oil,Squalene, Imidazolidinyl Urea and Sodium Ascorbyl Phosphate. A preferredembodiment of a lipid based colloidal carrier system is shown in Table2.

TABLE 2 Conc. range Quantity Ingredients (INCI) wt % wt % CAS-No.Phosphatylcholine 0-2-80 40.00 8002-43-5 Sphingomyeline 0-2-80 20.0085187-10-6 Linolenic Acid 0-5-50 10.00 463-40-1 Linoleic Acid 0-5-5010.00 60-33-3 Caprylic 0.2-40 10.00 73398-61-5 Triglyceride CapricTriglyceride 0.2-40 10.00 65381-09-1 Total: 100.00

In a further embodiment, the colloidal carrier system can also include apolycarbonate, a Polyvinylpyrrolidon (PVP), also Polyvidon orPovidonmembranes, preferably copovidone of a MW 10 nm-500 nm. Copovidonewill be used as film-forming agent and binder and also as carriersystem.

For use in the present invention, the lipophilic agents will beencapsulated within the bilayer system, whereas the hydrophilic agentswill be encapsulated in the aqueous phase of the system. Thus, thevitamin D precursor (such as 7-DHC or a derivative thereof) and thephospholipid(s) and optional additional lipophilic agents (e.g. retinylpalmitate and tocopheryl acetate) are directly incorporated in the oilphase of the colloidal carrier system at room temperature. In a separatestep, the HA and optional additional hydrophilic agents (e.g.riboflavin, niacinamide, dexpanthenol, folic acid, L-ascorbic acid) aremixed together separately in an aqueous solution. The aqueous solutioncomprising HA and optional additional hydrophilic agents are mixed tothe oil phase comprising the vitamin D precursor (such as 7-DHC or aderivative thereof). After emulsification of the two phases, thehydrophilic components (HA and additional hydrophilic agents) of thecompositions are present in aqueous compartments while the lipophiliccomponents of the compositions already insert themselves with the firststep in phospholipid bilayers of the particles.

Thus, in most preferred embodiments, the lipid based colloidal carriersystem (preferably lipid based vesicles such as liposomes, niosomes,tranferosomes) is composed of lecithin, linolenic acid, linoleic acid,phosphatidylcholin and paprylic/papric triglyceride, which is chargedwith the components Vitamin D3, such as 7-DHC or a derivative thereof,and HA and optionally at least one, preferably 1, 2, 3, 4, 5, 6 or 7 ofthe components retinyl palmitate, riboflavin, niacinamide, dexpanthenol,folic acid, L-ascorbic acid and tocopheryl acetate. The uniquely chargedand stable carrier system will allow penetrating the skin in the upperlayers of the skin to allow the synergistic compositions of theinvention to take effect directly at the desired site.

Depending on the nature and type of application, the compositions of theinvention may further comprise one or more pharmaceutically acceptableadditives, excipients, adjuvants commonly used in formulations used forapplication to the skin and/or mucous membranes.

Typical additives include e.g. a relevant UV filter system or one ormore UVA/B protectants for prevention of photodamage and sun UVA and UVBprotection such as Fillagrine trans-Urocanin Acide, ButylMethoxydibenzoylmethane Neo Heliopan 357 Eusolex 9020, Parsol 1789,Methylene Bis-Benzotriazolyl Tetramethylbutylphenol (nano), Tinosorb M,Ethylhexyl Triazone Uvinul T 150, Bis-Ethylhexyloxyphenol MethoxyphenylTriazine Tinosorb S, Ethylhexyl Methoxycinnamate Uvinul MC 80, ParsolMCX, Neo Heliopan AV 4, and the like. The UV filter(s) are embedded withthe active ingredients and auxiliary ingredients for the prevention ofskin erythema and actinic keratosis, or other forms of non melanoma skincancer (NMSC).

Typical adjuvants include e.g. surfactants, emulsifying agents,emollients, thickening agents, conditioning conservants, bufferingagents, humectants, perfuming agents, and the like. Thus, the carriersystem may further comprise one or more surfactants. The term“surfactant” refers to a material which lowers the surface tension of aliquid and the interfacial tension between two liquids, allowing theireasier spreading. Surfactants have a hydrophilic head that is attractedto water molecules and a hydrophobic tail that repels water andsimultaneously attaches itself to oil and grease in dirt. These opposingforces loosen the dirt and suspend it in the water, having the abilityto remove it from surfaces such as the human skin, textiles, and othersolids, when surfactants are dissolved in water. Examples of appropriatesurfactant agents include, but are not limited to, non-ionic, ionic(either anionic or cationic) or zwitterionic (or amphoteric wherein thehead of the surfactant contains two oppositely charged groups)surfactants. Examples of anionic surfactants include, but are notlimited to, those based on sulfate, sulfonate or carboxylate anions suchas perfluorooctanoate (PFOA or PFO), alkyl benzene sulfonate, soaps,fatty acid salts, or alkyl sulfate salts such asperfluorooctanesulfonate (PFOS), sodium dodecyl sulfate (SDS), ammoniumlauryl sulfate, or sodium lauryl ether sulfate (SLES). Examples ofcationic surfactants include, but are not limited to, those based onquaternary ammonium cations such as or alkyltrimethylammonium includingcetyl trimethylammonium bromide (CTAB) a.k.a., or hexadecyl trimethylammonium bromide, cetylpyridinium chloride (CPC), polyethoxylated tallowamine (POEA), benzalkonium chloride (BAC), or benzethonium chloride(BZT). Examples of zwitterionic surfactants include, but are not limitedto dodecyl betaine, cocamidopropyl betaine, or coco ampho glycinate.Examples of non-ionic surfactants include, but are not limited to, alkylpoly(ethylene oxide), alkylphenol poly(ethylene oxide), copolymers ofpoly(ethylene oxide), poly(propylene oxide) (commercially calledPoloxamers or Poloxamines), alkyl polyglucosides including octylglucoside and decyl maltoside, fatty alcohols including cetyl alcoholand oleyl alcohol, cocamide MEA, cocamide DEA, or polysorbates includingtween 20, tween 80, or dodecyl dimethylamine oxide. Preferably, thesurfactant is foaming and skin friendly, including polysorbates, such aspolysorbate 20 or 40, coco glucoside, lauryl glucoside, decyl glucoside,lauryl sulfates such as ammonium, sodium, magnesium, MEA, triethylamine(TEA), or mipa lauryl sulfate, cocamidopropyl betain, or sodium alkylsulfosuccinates.

In specific embodiments the surfactant is at least one polysorbate, e.g.polysorbate 10-150, which are non-ionic surfactants commonly used asexcipients and emulsifiers. Preferably the polysorbate is apolysorbate-type nonionic surfactant formed by the ethoxylation ofsorbitan before the addition of lauric acid as Scattics, PS20 as AlkestTW 20 and Tween 20. Polysorbates have efficiency in the stabilization ofthe colloidal carrier and in the presence of liquid lipids withdifferent fatty acid C-chains produces with less organized crystallinestructure can provides better loading capacity for active substanceaccommodation. The effect of polysorbate will be in the stabilizationthe carrier system through the physiochemical properties of theformulated nanoparticles. The colloidal carrier system are stabilizedwith polysorbate like polysorbate 20 or polysorbate 80. Polysorbate willbe used as better dispersing agent for the liposomal carrier system. Thesmall size and superior particle surface to volume ratio would increaseloading efficiency and bioavailability of the active substance, thusmaking the liposomal carrier system a more efficient delivery system.

In other specific embodiments the surfactant is polyvinylpyrrolidone(PVP), which is known to either prevent precipitation or reduce the sizeof the resulting particles of the active ingredients or auxiliarysubstances with strongly pH-dependent aqueous solubility. The PVP likepoloxamer/copovidone will be used to stabilize particles in theliposomal formulation. It is presumed, that the dissolution efficiencyis higher with Polyvinylpyrrolidone (PVP) and is increased withincreased polymer concentration. PVP is typically used as stabilisationand to increase efficiency and bioavailability of the liposomal carriersystem.

Thus, in specific embodiments the carrier system may further compromisea polycarbonate, Polyvinylpyrrolidon (PVP), Polyvidon, Povidonmembranes,Povidone, Copovidone, Hypromellose and Eudragit EPO, preferablyCopovidone of a MW 10 nm-500 nm.

The amount of the surfactant in the compositions of the presentinvention is between 0.5 and 10 wt % of the total weight of thecomposition according to the present invention.

The term “emollient” agent refers to an agent that softens and soothesthe skin in order to correct dryness and scaling of the skin,lubricating the skin surface, encouraging skin water retention, andaltering product textures. Examples of appropriate topical emollientagents include, but are not limited to, octyl hydroxystearate, lanolin,caprylic/capric triglyceride, cetyl palmitate, octyldodecanol, cetylalcohol, isopropyl isostearate, glyceryl dilaurate, isopropyl myristate,palm alcohol, dimethicone, squalane, plukenetia volubilis seed oil,butyrospermum parkii butter, sucrose cocoate, or their mixtures.Preferably the emollient is selected from the group consisting ofdimethicone, squalane, plukenetia volubilis seed oil, butyrospermumparkii butter, caprylic/capric triglyceride, octyldodecanol, or theirmixtures. The amount of emollient agent in the compositions of thepresent invention is between 10 and 30 wt % of the total weight of thecomposition according to the present invention.

The term “humectant” agent refers to a hygroscopic agent which attractswater molecules from the surrounding environment though eitherabsorption or adsorption, preventing the skin from losing moisture.Examples of appropriate topical humectants include, but are not limitedto, glycerin, diglycerin, ethylhexylglycerin, glucose, honey, lacticacid, polyethylene glycol, propylene glycol, sorbitol, sucrose, orthrealose. Preferably, the humectant is selected group consisting ofglycerin, diglycerin, ethylhexylglycerin, and their mixtures. The amountof the humectants in the compositions of the present invention isbetween 0.5-15 wt %, preferably 0.5-10 wt %, of the total weight of thecomposition according to the present invention.

The term “thickening agent” or “thickener” or “viscosity agent” which isherein used interchangeably refers to a material that increases itsviscosity without substantially modifying its other properties. Examplesof appropriate viscosity agents include, but are not limited to,cellulose or their derivatives such as hydroxypropyl methylcellulose,polyethylene glycol, microcrystalline cellulose, cetearyl alcohol,alginates, branched polysaccharides, fumed silica, xanthan gum,carbomer, and polyacrylates. Preferably, the viscosity agent is selectedgroup consisting of microcrystalline cellulose, cetearyl alcohol,cellulose, xanthan gum, and carbomer. The amount of the viscosity agentsin the compositions of the present invention is between 0.5 and 15 wt %,preferably 0.5-10 wt %, of the total weight of the composition accordingto the present invention.

The term “emulsifying agent” or “emulsifier” which is herein usedinterchangeably refers to a material that reduces surface tension,promoting the formation of intimate mixtures of non-miscible liquids byaltering the interfacial tension. Emulsifier stabilizes an emulsion byincreasing its kinetic stability. Examples of appropriate emulsifierinclude, but are not limited to, glyceryl trioleate, glyceryl oleate,acetylated sucrose distearate, sorbitan trioleate, polyoxyethylenemonostearate, glycerol monooleate, sucrose distearate, polyethyleneglycol monostearate, octyl phenoxypoly (ethyleneoxy) ethanol, deacylerinpenta-isostearate, sorbitan sesquioleate, hydroxylated lanolin,lecithin, lanolin, triglyceryl diisostearate, polyoxyethylene oleylether, calcium stearoyl-2-lactylate, sodium lauroyl lactylate, sodiumstearoyl lactylate, cetearyl glucoside, methyl glucoside sesquistearate,sorbitan monopalmitate, methoxy polyethylene glycol-22/dodecyl glycolcopolymer, polyethylene glycol-45/dodecyl glycol copolymer, polyethyleneglycol 400 distearate and glyceryl stearate, candelilla/jojoba/rice branpolyglyceryl-3 esters, cetyl phosphate, potassium cetyl phosphate, ortheir mixtures. Preferably, the emulsifier is selected group consistingof glyceryl oleate, lecithin, sodium lauroyl lactylate, sodium stearoyllactylate, glyceryl stearate, candelilla/jojoba/rice bran polyglyceryl-3esters, and their mixtures. The amount of the emulsifier in thecompositions of the present invention is between 0.5 and 15 wt %,preferably 0.5-10 wt %, of the total weight of the composition accordingto the present invention.

The term “pH-regulating” or “buffering” agent refers to acids or basesthat can be used to adjust the pH of the finished product to the desiredlevel, without affecting the stability of the solution. Examples ofappropriate topical pH-regulating agents include, but are not limitedto, acetic acid, lactic acid, citric acid, gluconic acid, ethanolamine,formic acid, oxalic acid, tartaric acid, potassium hydroxide, sodiumhydroxide, triethanolamine, or their mixtures. Preferably, thepH-regulating agent is selected group consisting of triethanolamine,sodium hydroxide, lactic acid, and citric acid. The amount of thepH-regulating agent in the compositions of the present invention isbetween 0.01 and 1 wt % of the total weight of the composition accordingto the present invention.

The term “conditioning conservant” refers to a compound that has amoisturizing function, more specifically a compound that acts on thebarrier function, for the purpose of keeping the stratum corneummoisturized, such as ceramides, sphingoid-based compounds, lecithins,glycosphingolipids, phospholipids, cholesterol and its derivatives,phytosterols (stigmasterol, β-sitosterol or campesterol), essentialfatty acids, 1,2-diacylglycerol, 4-chromanone, pentacyclic triterpenes,petroleum jelly and lanolin; or a compound which directly increases thewater content of the stratum corneum, such as threalose and itsderivatives, glycerol, pentanediol, pidolates, serine, xylitol,peroxyethanol, sodium lactate, glyceryl polyacrylate, ectoin and itsderivatives, chitosan, oligo- and polysaccharides, cyclic carbonates,N-lauroylpyrrolidonecarboxylic acid and N-α-benzoyl-L-arginine. Theamount of duch compounds in the composition of the present invention isfrom 0.001 wt % to 30 wt %, preferably from 0.01 to 20 wt %, of thetotal weight of the composition according to the present invention.

The term “perfuming agent” refers to any perfume or aroma which iscapable of releasing an agreeable odor. The perfuming substancecontained in the compositions of the invention may derive from perfumesand aromas of natural or synthetic origin and mixtures thereof. Examplesof perfumes and aromas of natural origin are flower extracts (lily,lavender, rose, jasmine, ylang-ylang), stems and leaves (patchouli,geranium, bitter leaf), fruits (coriander, anis, cumin, juniper), fruitskin (bergamot, lemon, orange), roots (angelica, celery, cardamom, iris,sweet flag), wood (pinewood, sandalwood, lignum vitae, pink cedar),herbs and graminaceae (tarragon, lemon grass, sage, thyme), needles andbranches (spruce, fir, pine, dwarf pine), resins and balsams (galbanum,gum elemi, gum benzoin, myrrh, frankincense, opopanax).

Preferably, the quantity of perfuming agents is from 1 wt % to 30 wt %by weight, more preferably 2 wt % to 25 wt % by weight with respect tothe total composition weight. A preferred composition is shown in Table3.

TABLE 3 Ingredients (INCI) without active substances and phospholipidesof the Quantity carrier system wt % Function CAS-No. Aqua destillata56.79 Solvent 7732-18-5 Isopropyl Myristate 1.0 Emulsifying Emolient110-27-0 Palmitoyl Tripeptide-5 1.0 Emulsifying Emolient 623172-56-5 95Palmitic Acid 1.0 Emulsifying Emolient 57-10-3 Glycerin 3.0 EmulsifyingEmolient 56-81-5 Lupinus albus seed oil 2.0 Emulsifying Emolient545-47-1 Palmitoyl-Pentapeptid 1.0 Emulsifying Emolient 214047-00-4Cetearyl Alcohol 1.0 Emulsifying Emolient 67762-27-0/ 8005-44-5Polidocanol 3.0 Emulsifying Emolient 3055-99-0 Squalene 1.0 EmulsifyingEmolient 111-02-4 Hydroxypalmitoyl 1.0 Emulsifying Emolient 190249-36-6Sphinganine Imidazolidinyl Urea 2.0 Humectant 39236-46-9 Prunus amygalusdulcis 5.0 Conditioning 8007-69-0/ oil Conservant 90320-37-9 Pyridoxine1.0 Conditioning 58-56-0 hydrochloride Conservant Sodium Ascorbyl 2.0Conditioning 66170-10-3 Phosphate Conservant Phenoxyethanol/ 1.0Conditioning 122-99-6 Peroxyethanol Conservant Green Tea Polyphenole 1.0Conditioning 84650-60-2 Conservant Sodium Citrate 1.0 Buffering 68-04-2PEG-5 Glyceryl 1.0 Surfactant 51158-08- Stearate 8 138860-92-1 StearicAcid 1.0 Surfactant 57-11-44 Fillagrine trans- 1.0 UV-filter104-98-3-3465-72-3 Urocanin Acide Butyl 4.8 UV Filter 70356-09-1Methoxydibenzoylmethane Methylene Bis- 3.5 UV Filter 103597-45-1Benzotriazolyl Tetramethylbutylphenol (nano) Ethylhexyl Triazone 2.0 UVFilter 88122-99-0 Bis- 0.8 UV Filter 187393-00-6 EthylhexyloxyphenolMethoxyphenyl Triazine Ethylhexyl 0.1 UV Filter 5466-77-3Methoxycinnamate Octyldodecanol 1.0 Perfuming 5333-42-6 GeraniumMaculatum 0.002 Perfuming 84650-10-2 Oil Citrus Aurantium 0.002Perfuming 8008-57-9 Dulcis Oil Citrus Medica 0.002 Perfuming 8008-56-8/Limonum Peel Oil 84929-31-7 Aniba Rosaeodora Oil 0.004 Perfuming83863-32-5 Total: 100.00

Lipid based colloidal carrier system (e.g. lipid based vesicles such asliposomes, niosomes, tranferosomes) can be prepared by any of thetechniques known (for the preparation of lipid based carrier systems ingeneral, see e.g. Liposomes, eds. Angel Catala, pub. InTech, 2017 (ISBN978-953-51-3580-7), or of liposomal carriers see e.g. Liposomes, Methodsand Protocols, Springer Protocols, eds. D'Souza, Gerard G. M., 2017).For example, the colloid can be formed by any conventional technique forpreparing multilamellar lipid vesicles (MLVs), that is, by placing thelipophilic vitamin D3 or precursor thereof with one or more lipids in asuitable vessel, dissolving the lipids in an organic solvent, e.g.chloroform, and evaporating the organic solvent to obtain a lipid film.In a subsequent step hydration of the lipid film is achieved by addingan aqueous solution containing the hydrophilic components including thehyaluronic acid. Typically the obtained lipid suspension is subjected toswirling or vortexing to give the final composition according to theinvention. Alternatively, techniques used for producing largeunilamellar lipid vesicles (LUVs), such as reverse-phase evaporation,infusion procedures, and detergent dilution, can be used to produce theliposomes. A review of these and other methods for producing lipidvesicles can be found in the text Liposome Technology, Volume I, GregoryGregoriadis Ed., CRC Press, Boca Raton, Fla., (1984), which isincorporated herein by reference. For example, the lipid-containingparticles can be in the form of steroidal lipid vesicles, stableplurilamellar lipid vesicles (SPLVs), monophasic vesicles (MPVs), orlipid matrix carriers (LMCs). In the case of MLVs, if desired, theliposomes can be subjected to multiple (five or more) freeze-thaw cyclesto enhance their trapped volumes and trapping efficiencies and toprovide a more uniform interlamellar distribution of solute.

In one embodiment, the liposomes are for example prepared by hot highpressure homogenization to reach high encapsulation efficiency (EE). Theencapsulation efficiency will give the percentage of active substancethat is successfully entrapped/adsorbed into nanoparticles and will becarried into the depper layers of the skin. A major obstacle to theapplication of nanostructured lipid carriers (NLCs) as carriers forhydrophilic active substances is the limited loading capacity (LC) andencapsulation efficiency (EE) of NLCs for these molecules, with wt % EEbeing equal to the [(active substance added−Free “unentrapped activesubstance”)/active substance added]*100 (thus as an example, an wt % EEof 5 wt % means that 5 wt % of the active substance is entrapped intothe carrier system).

The phase transfer temperature from the gel form to the crystalline twodimensional grid states with less mobility in a fluid crystallinestructure. The phase transfer temperature of the mentioned lipids isdepending of the head group, chain lengths and the saturation levelesters of fatty acids. The temperature will be from −20° C. to 60° C.and can be established with the thermoanalytic methods. Embedded in thefluid crystalline phase the mobility of the lipophilic agents increasesand can exchange the place within the lipid layers, but not abandon thelipid layers [46].

The physical structure of the multilamellar layer system will be createdthroughout interactions between the phospholipids and the aqueous mediumwith the high pressure homogenisation and dehydration of dry lipids.With this method multilamellar vessels (MLV) are built. Thepolycarbonate membranes of a size 10-500 nm will be used for theliposomes extrusion. The homogenisation and size will be determinedthroughout the pore diameter of the filter and the number of theextrusion steps. Aim will be to reach the highest encapsulationefficiency.

The use of this carrier system has unique physicochemical properties,such as ultra-small size (small particles from 1-100 nm dimensionrange), large surface area to mass ratio, and high reactivity, which aredifferent from bulk materials of the same composition. These propertiesare being used to overcome the limitation of skin penetration withlarger size of molecules and encapsulate as needed lipophilic andhydrophilic substances to pass the skin barrier.

In a further aspect the invention is directed towards suitableformulations of the compositions of the invention for topical ortransdermal application.

The compositions of this invention can be used in different types oftopical or transdermal applications, which may be in solid, liquid orsemisolid form. Thus, suitable formulations include, but are not limitedto, emulsions (e.g. oil and/or silicone in water emulsions, water-in-oiland/or silicone emulsions, water/oil/water or water/silicone/water typeemulsions, and oil/water/oil or silicone/water/silicone type emulsions),microemulsions, aqueous dispersions, oils, milks, balsams, foams,aqueous or oily lotions, aqueous or oily gels, creams, solutions,hydroalcoholic solutions, hydroglycolic solutions, hydrogels, serums,ointments, mousses, pastes, sprays or aerosols, as well as inclusion ofthe compositions of the invention in any transdermal patches. In atypical transdermal therapeutic system, such as a patch or pad, thecompositions of the invention (with or without at least one auxiliaryagent) are embedded, if desired, in combination with penetrationreinforcing agents and/or crystallisation inhibitors. Thus, in specificembodiments the compositions are in form of a cream or a gel or alotion, in other specific embodiments the compositions are in form oftransdermal therapeutic system, such as a patch or pad.

In a further aspect, the invention is directed towards the use of thecomposition (and formulations thereof) of the invention in theprevention and/or treatment of skin photo damage symptoms, in particularin the prevention and/or treatment of skin atrophy, skin dyspigmentation(patches/spots), photodermatitis (erythema: skin inflammation andredness), telangiectasia, (couperose) and prevention of actinickeratosis, as well to protect the skin from the sun, UVA and UVBradiation.

Thus, the present invention contemplates a method of prevention and/ortreatment of photodamage of the skin of a subject comprisingadministering a composition (and topical formulations thereof) of theinvention to the subject in an amount effective to stop the photodamageprocess, i.e. to inhibit reactive oxygen species ROS, hyperoxide O-2 andnitric oxide (NO), and therefore accumulation of cytotoxic peroxynitrite(ONOO—).

The compositions (or formulations thereof) may be either administered atregular intervals as needed (e.g., once, twice or several times a day)or in an essentially continuous manner (e.g. via a transdermal patch).

The following examples are representative examples to illustrate theinvention, without limiting the scope of the invention.

EXAMPLES Methods and Materials

Dried phospholipides were dispersed at room temperature in aqueoussolution forming spontaneously spheric colloids. 7-DHC of purity 98.7%(hplc; area %) was liquefied at temperatures between 140 and 150° C.Propylenglycol was added to the liquefied 7-DHC and the obtained mixturewas admixed to the phospholipids colloids under vigorous stirring atroom temperature to obtain colloid forming spheres of 20-150 nm adding7-DHC in the oil phase of the colloids. HA (2% low molecular 4 KDa 96.8%of purity, 0.5 wt % of medium molecular 48.3 KDa 97.3% purity and 0.5 wt% of high molecular 1.78×10⁶ Da eye drop grade of purity 100%) wasdispersed/dissolved under stirring in water and added under stirring atroom temperature separately. HA mixtures were added to the phospholipidscolloids already charged with 7-DHC and stirred for 20 minutes at roomtemperature to obtain spontaneous formation of a homogeneous hydrogel.The microscopic analysis showed spheric particles of 20-150 nm size. Thehydrogel was macroscopic and according to HPLC analysis stable showingthe same concentration of added 7-DHC over 6 months.

For the lipid phase 7-DHC of purity 98.7% (hplc; area %) was used(liquefied at temperatures between 140 and 150° C.). Propylenglycol wasused as the organic solvent for the 7-DHGC and the phospholipids.

HA (in form of a mixture of 2% low molecular 4 KDa 96.8% of purity, 0.5wt % of medium molecular 48.3 KDa 97.3% purity and 0.5 wt % of highmolecular 1.78×10⁶ Da eye drop grade of purity 100%) was used for theaqueous phase.

Stirring for 20 minutes at room temperature resulted in a spontaneousformation of a homogeneous hydrogel. The microscopic analysis showedspheric particles of 20-150 nm size.

Stability studies showed high stability (>98%) for 6 months and constantcontent of 1.5% of DHC in the analytic (HPLC).

Example 1 without UV-Filter System Cream

TABLE 4 Quantity Ingredients (INCI) wt % CAS-No. 7-DHC 0.15 434-16-2 HA3 9004-61-9 Retinyl Palmitate 0.5 79-81-2 Skin Conditioning Riboflavin0.1 83-88-5 Niacinamide 4.0 98-92-0 Dexpanthenol 2.5 81-13-0 Folic acid0.05 59-30-3 L-ascorbic acid 3.0 50-81-7 Tocopheryl Acetate 2.07695-91-2 Aqua destillata 46.196 7732-18-5 Solvent PEG-5 GlycerylStearate 1.0 51158-08-8 Surfactant 138860-92-1 Stearic Acid 1.0 57-11-44Emulsifying Emolient Isopropyl Myristate 1.0 110-27-0 Palmitic Acid 1.057-10-3 Lupinus albus Ölextrakt 2.0 545-47-1 Palmitoyl-Pentapeptid 1.0214047-00-4 Prunus amygalus dulcis oil 5.0 8007-69-0/ 90320-37-9Squalene 1.0 111-02-4 Polidocanol 3.0 3055-99-0Hydroxypalmitoylsphinganin 1.0 190249-36-6 Pyridoxine HCL 5.0 8007-69-0/Conditioning 90320-37-9 Conservant Sodium Ascorbyl Phosphate 2.066170-10-3 Sodium Citrate 1.0 6132-04-3 Buffer Lecithin 2 8002-43-5Carrier encapsulation formulary* Sphingomyeline 2 85187-10-6 LinolenicAcid 1 463-40-1 Linoleic Acid 1 60-33-3 Phosphatidylcholin 3 26853-31-6Caprylic Triglyceride 1 73398-61-5 Octyldodecanol 1.0 5333-42-6 PerfumeCitrus Aurantium Dulcis Oil 0.002 8008-57-9 Citrus Medica Limonum 0.0028008-56-8/ Peel Oil 84929-31-7 VP/Eicosene Copolymer 2.5 28211-18-9 FilmFormer Total: 100

Example 2 without UV-Filter System Gel

TABLE 5 Quantity Ingredients (INCI) wt % Function CAS-No. 7-DHC 0.15Active ingredient 434-16-2 ROS binder HA 3 Active ingredient 9004-61-9Humectant Retinyl Palmitate 0.5 Skin Conditioning 79-81-2 Riboflavin 0.1Skin Conditioning 83-88-5 Niacinamide 4.0 Skin Conditioning 98-92-0Dexpanthenol 2.5 Skin Conditioning 81-13-0 Folic acid 0.05 SkinConditioning 59-30-3 L-ascorbic acid 3.0 Skin Conditioning 50-81-7Tocopheryl 2.0 Skin Conditioning 7695-91-2 Acetate Aqua destillata 65.77Solvent 7732-18-5 Stearic Acid 1.0 Surfactant 57-11-44 Isopropyl 1.0Emulsifying Emolient 110-27-0 Myristate Palmitic Acid 1.0 EmulsifyingEmolient 57-10-3 Cetearyl Alcohol 1.0 Emulsifying Emolient 67762-27-0/8005-44-5 Polidocanol 2.0 Emulsifying Emolient 3055-99-0 Sodium Citrate1.0 Buffering 6132-04-3 Lecithin 1 Carrier encapsulation 8002-43-5formulary* Sphingomyeline 2 Carrier encapsulation 85187-10-6 formulary*Linolenic Acid 1 Carrier encapsulation 463-40-1 formulary* Linoleic Acid1 Carrier encapsulation 60-33-3 formulary* Phosphatidylcholin 2.0Carrier encapsulation 26853-31-6 formulary* Caprylic 1 Carrierencapsulation 73398-61-5 Triglyceride formulary* Capric 1 Carrierencapsulation 65381-09-1 Triglyceride formulary* Triethanolamine 0.08ph-adjusting 102-71-6 Peroxyethanol 3 Conservant 95684-29-0 Total: 100

Example 3 without UV-Filter System Serum

TABLE 6 Quantity Ingredients (INCI) wt % Function CAS-No. 7-DHC 0.15Active 434-16-2 ingredient ROS binder HA 3 Active 9004-61-9 ingredientHumectant Dexpanthenol 2.5 Skin 81-13-0 Conditioning Folic acid 0.05Skin 59-30-3 Conditioning Tocopheryl Acetate 2.0 Skin 7695-91-2Conditioning Aqua destillata 70.3 Solvent 7732-18-5 PEG-5 GlycerylStearate 1.0 Surfactant 51158-08-8 138860-92-1 Stearic Acid 1.0Surfactant 57-11-44 Isopropyl Myristate 1.0 Emulsifying 110-27-0Emolient Palmitic Acid 1.0 Emulsifying 57-10-3 EmolientPalmitoyl-Pentapeptid 1.0 Emulsifying 214047-00-4 Emolient Polidocanol2.0 Emulsifying 3055-99-0 Emolient Hydroxypalmitoylsphinganin 1.0Emulsifying 190249-36-6 Emolient Sodium Citrate 1.0 Buffering 6132-04-3Lecithin 1 Carrier 8002-43-5 encapsulation formulary* Sphingomyeline 2Carrier 85187-10-6 encapsulation formulary* Linolenic Acid 1 Carrier463-40-1 encapsulation formulary* Linoleic Acid 1 Carrier 60-33-3encapsulation formulary* Phosphatidylcholin 3 Carrier 26853-31-6encapsulation formulary* Caprylic Triglyceride 1 Carrier 73398-61-5encapsulation formulary* Capric Triglyceride 1 Carrier 65381-09-1encapsulation formulary* Peroxyethanol 3 Conservant 95684-29-0 Total:100

Example 4 with UV-Filter System Face

TABLE 7 Quantity Ingredients (INCI) wt % Function CAS-No. 7-DHC 0.15Active ingredient 434-16-2 ROS binder HA 3 Active ingredient 9004-61-9Humectant Aqua destillata 55.15 Solvent 7732-18-5 PEG-5 GlycerylStearate 1.0 Surfactant 51158-08-8 138860- 92-1 Stearic Acid 1.0Surfactant 57-11-44 Isopropyl Myristate 1.0 Emulsifying 110-27-0Emolient Palmitic Acid 1.0 Emulsifying 57-10-3 Emolient Pyridoxine HCL5.0 Conditioning 8007-69-0/90320-37-9 Conservant Sodium AscorbylPhosphate 2.0 Conditioning 66170-10-3 Conservant Polidocanol 3.0Emulsifying 3055-99-0 Emolient Hydroxypalmitoylsphinganin 1.0Emulsifying 190249-36-6 Emolient Sodium Citrate 1.0 Buffering 6132-04-3Lecithin 1 Carrier 8002-43-5 encapsulation formulary* Sphingomyeline 2Carrier 85187-10-6 encapsulation formulary* Linolenic Acid 1 Carrier463-40-1 encapsulation formulary* Linoleic Acid 1 Carrier 60-33-3encapsulation formulary* Phosphatidylcholin 3 Carrier 26853-31-6encapsulation formulary* Caprylic Triglyceride 1 Carrier 73398-61-5encapsulation formulary* Capric Triglyceride 1 Carrier 65381-09-1encapsulation formulary* VP/Eicosene Copolymer 2.5 Film forming28211-18-9 Fillagrine trans-Urocanin 1 UV-filter 104-98-3-3465-72-3Acide Butyl 3.8 UV Filter 70356-09-1 Methoxydibenzoylmethane NeoHeliopan 357, Eusolex 9020, Parsol 1789 Methylene Bis- 2.5 UV Filter103597-45-1 Benzotriazolyl Tetramethylbutylphenol (nano) Tinosorb MEthylhexyl Triazone Uvinul 2.0 UV Filter 88122-99-0 T 150Bis-Ethylhexyloxyphenol 0.8 UV Filter 187393-00-6 Methoxyphenyl TriazineTinosorb S Ethylhexyl 0.1 UV Filter 5466-77-3 Methoxycinnamate Uvinul MC80, Parsol MCX, Neo Heliopan AV 4 Peroxyethanol 3 Conservant 95684-29-0Total: 100,000

Example 5 with UV-Filter System Body

TABLE 8 Quantity Ingredients (INCI) wt % Function CAS-No. 7-DHC 0.15Active ingredient 434-16-2 ROS binder HA* 3 Active ingredient 9004-61-9Humectant Retinyl Palmitate 0.2 Skin 79-81-2 Conditioning Riboflavin 0.1Skin 83-88-5 Conditioning Niacinamide 2.0 Skin 98-92-0 ConditioningDexpanthenol 2.5 Skin 81-13-0 Conditioning Folic acid 0.05 Skin 59-30-3Conditioning L-ascorbic acid 2.0 Skin 50-81-7 Conditioning TocopherylAcetate 1.0 Skin 7695-91-2 Conditioning Aqua destillata 56.263 Solvent7732-18-5 Stearic Acid 1.0 Surfactant 57-11-44 Pyridoxine HCL 3.0Conditioning 8007-69-0/90320-37-9 Conservant Sodium Ascorbyl 2.0Conditioning 66170-10-3 Phosphate Conservant Polidocanol 3.0 Emulsifying3055-99-0 Emolient Sodium Citrate 1.0 Buffering 6132-04-3 Lecithin 1Carrier 8002-43-5 encapsulation formulary* Sphingomyeline 2 Carrier85187-10-6 encapsulation formulary* Linolenic Acid 1 Carrier 463-40-1encapsulation formulary* Linoleic Acid 1 Carrier 60-33-3 encapsulationformulary* Phosphatidylcholin 3 Carrier 26853-31-6 encapsulationformulary* Caprylic Triglyceride 1 Carrier 73398-61-5 encapsulationformulary* Capric Triglyceride 1 Carrier 65381-09-1 encapsulationformulary* VP/Eicosene Copolymer 2.5 Film forming 28211-18-9 Trisodium0.037 Chelating 20846-91-7/178949- Ethylenediamine 82-1 DisuccinateTrisodium Ethylenediamine Disuccinate Solution, Natrlquest E30 3)Fillagrine trans-Urocanin 1 UV-filter 104-98-3-3465-72-3 Acide Butyl 3.8UV Filter 70356-09-1 Methoxydibenzoylmethane Neo Heliopan 357, Eusolex9020, Parsol 1789 Methylene Bis- 2.5 UV Filter 103597-45-1Benzotriazolyl Tetramethylbutylphenol (nano) Tinosorb M EthylhexylTriazone 2.0 UV Filter 88122-99-0 Uvinul T 150 Bis-Ethylhexyloxyphenol0.8 UV Filter 187393-00-6 Methoxyphenyl Triazine Tinosorb S Ethylhexyl0.1 UV Filter 5466-77-3 Methoxycinnamate Uvinul MC 80, Parsol MCX, NeoHeliopan AV 4 Total: 100,000

1. A composition comprising at least one vitamin D3 or precursor orderivative thereof and at least one hyaluronic acid or derivativethereof encapsulated in a lipid based colloidal carrier system.
 2. Thecomposition according to claim 1 wherein the vitamin D3 precursor is7-DHC or a derivative thereof.
 3. The composition according to claim 1wherein the hyaluronic acid is a mixture of low molecular, mediummolecular and high molecular weight hyaluronic acid.
 4. The compositionaccording to claim 1 wherein the lipid based colloidal carrier systemincludes liposomes, niosomes, transferosomes, micelles, nanoparticles,microemulsions and others, preferably liposomes, niosomes,transferosomes, most preferably liposomes.
 5. The composition accordingto claim 1 further comprising one or more further components selectedfrom a vitamin A, preferably retinyl palmitate, at least one vitamin B,a vitamin C, preferably L-ascorbic acid and a vitamin E, preferablytocopheryl acetate.
 6. The composition according to claim 5 wherein theat least one vitamin B is riboflavin (vitamin B2), niacinamide (vitaminB3), dexpanthenol (Provitamin B5), and/or folic acid (vitamin B9). 7.The composition according to claim 1 further comprising i) retinylpalmitate (vitamin A), (ii) riboflavin (vitamin B2), (iii) niacinamide(vitamin B3), (iv) dexpanthenol (Provitamin B5), (v) folic acid (vitaminB9), (vi) L-ascorbic acid (vitamin C) and (vii) tocopheryl acetate(vitamin E).
 8. The composition according to claim 1 further comprisingat least one UV-filter.
 9. The composition according to claim 1 furthercomprising at least one adjuvant selected from surfactants, emulsifyingagents, emollients, thickening agents, conditioning conservants,buffering agents, humectants, perfuming agents.
 10. The compositionaccording to claim 9 wherein the surfactant is a polycarbonate,polyvinylpyrrolidon (PVP), polyvidon, ovidonmembranes, povidone,copovidone, hypromellose or Eudragit EPO.
 11. The composition accordingto claim 9 wherein the surfactant is a polysorbate 10-150, preferablypolysorbate PS20 as Alkest TW 20 and Tween
 20. 12. The compositionaccording to claim 1 wherein the lipid based colloidal carrier systemcomprises one or more components selected from phospholipids, lecithin,sphingomyelin, linolenic acid, linoleic acid, phosphatidylcholine andcaprylic/capric triglyceride.
 13. A topical formulation comprising acomposition according to claim 1, liquid or semisolid form, preferablyin form of emulsions, microemulsions, aqueous dispersions, oils, milks,balsams, foams, aqueous or oily lotions, aqueous or oily gels, creams,solutions, hydroalcoholic solutions, hydroglycolic solutions, hydrogels,serums, ointments, mousses, pastes, sprays or aerosols, or transdermalpatches.
 14. A use of a composition according to claim 1 in theprevention and/or treatment of skin disorders, inflamed skin, as eczema,rosacea, atopic dermatitis, psoriasis, including photodamage (such assun-induced inflamed and reddened skin) skin atrophy, skindyspigmentation (patches/spots), photodermatitis (erythema: infalmmatedand reddened skin), telangiectasia, (couperose) and prevention ofphotodermatitis (erythema), actinic keratosis.
 15. A method ofprevention and/or treatment of photodamage of the skin of a subjectcomprising administering a composition according to claim 1 to thesubject in an amount effective to prevent and/or treat photodamage (suchas sun-induced damage), in particular in the prevention and/or treatmentof skin atrophy, skin dyspigmentation (patches/spots), photodermatitis(erythema: inflamed and reddened skin), telangiectasia, (couperose) andprevention of photodermatitis (erythema), actinic keratosis.